生物学杂志 ›› 2022, Vol. 39 ›› Issue (5): 32-.doi: 10.3969/j.issn.2095-1736.2022.05.032

• 研究报告 • 上一篇    下一篇

棉花GhVTC2基因促进烟草BY2悬浮细胞和拟南芥主根伸长

  

  1. 1. 石河子大学 生命科学学院, 石河子 832003; 2. 绿洲城镇与山盆系统生态兵团重点实验室,
    石河子 832003; 3. 新疆植物药资源利用教育部重点实验室, 石河子 832003
  • 出版日期:2022-10-18 发布日期:2022-10-18
  • 通讯作者: 王斐,博士研究生,讲师,研究方向为植物分子生物学与基因工程,E-mail: feiw@shzu.edu.cn
  • 作者简介:董祥雨,硕士研究生,研究方向为生物化学与分子生物学,E-mail: 2982315787@qq.com
  • 基金资助:
    国家自然科学基金项目(31960413);石河子大学科技计划项目(CXPY202017,RCZK201905)

GhVTC2 gene promotes elongation of tobacco BY2 suspension cells and Arabidopsis main roots

  1. 1. College of Life Sciences, Shihezi University, Shihezi 832003, China; 2. Key Laboratory of Oasis Town and
    Mountain-basin System Ecology of Xinjiang Production and Construction Corps, Shihezi University, Shihezi 832003,
    China; 3. Key Laboratory of Xinjiang Phytomedicine Resource and Utilization of Ministry of Education,
    Shihezi University, Shihezi 832003, China
  • Online:2022-10-18 Published:2022-10-18

摘要: 为研究VTC2基因在棉花纤维发育及细胞伸长过程中的功能,从棉花中鉴定获得一个GhVTC2基因,表达分析表明:GhVTC2在棉纤维快速伸长发育时期显著累积,暗示其在细胞生长发育过程中的重要作用。构建植物过量表达载体35S:GhVTC2并遗传转化烟草BY2悬浮细胞和拟南芥,转基因烟草悬浮细胞获得显著的伸长;与野生型拟南芥相比,转基因拟南芥的主根伸长发育受到显著促进;转基因烟草细胞和拟南芥根中的AsA含量显著增加。通过克隆获得GhVTC2基因的启动子,序列分析显示,该启动子中包括多个乙烯响应元件ERE。结合启动子所包含ERE元件构建了启动子不同序列的5′缺失载体,乙烯前体1-氨基环丙烷-1-羧酸(ACC)处理转化不同缺失载体的烟草叶片的结果显示,ACC显著促进包含有ERE元件启动子的活性。研究结果为研究VTC2基因影响细胞伸长发育的功能及调控机制提供有效参考。

关键词: 棉花纤维, 抗坏血酸, 乙烯, 细胞伸长, 烟草悬浮细胞, 拟南芥

Abstract: In order to study the function of VTC2 gene in cotton fiber development and cell elongation, a GhVTC2 gene was identified from fast elongating fibers of cotton. Expression analysis showed that GhVTC2 accumulated significantly during the rapid elongation period of cotton fibers, suggesting its important role in cell growth and development. The plant overexpression vector 35S:GhVTC2 was constructed and transformed into tobacco BY2 suspension cells and Arabidopsis thaliana; compared with that of the non-transformed Arabidopsis plants and tobacco cells, the cell length was increased significantly in transgenic tobacco cells, and the elongation of main roots in transgenic Arabidopsis was significantly promoted. The contents of AsA in transgenic tobacco cells and Arabidopsis root were increased. The promoter of GhVTC2 gene was obtained by cloning, and sequence analysis showed that the promoter contained multiple ethylene responsive elements ERE. The 5′ deletion vectors of different promoter sequences with or without the ERE element were constructed and transformed into tobacco leaves treated by the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC). The results showed that ACC significantly promoted the activity of ERE-containing promoters. An effective reference is provided for studying the function and regulatory mechanism of GhVTC2 gene in cell elongation and development.

Key words: cotton fiber, ascorbic acid, ethylene, cell elongation, tobacco suspension cell, Arabidopsis

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